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Acidic lipase L ip I .3 from a P seudomonas fluorescens ‐like strain displays unusual properties and shows activity on secondary alcohols
Author(s) -
Panizza P.,
Syfantou N.,
Pastor F.I.J.,
Rodríguez S.,
Díaz P.
Publication year - 2013
Publication title -
journal of applied microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.889
H-Index - 156
eISSN - 1365-2672
pISSN - 1364-5072
DOI - 10.1111/jam.12089
Subject(s) - pseudomonas fluorescens , lipase , strain (injury) , pseudomonas , food science , pseudomonadaceae , triacylglycerol lipase , chemistry , pseudomonadales , microbiology and biotechnology , biology , enzyme , biochemistry , bacteria , genetics , anatomy
Aims Identification, cloning, expression and characterization of a novel lipase – L ip I .3 – from strain P seudomonas CR ‐611. Methods and Results The corresponding gene was identified and isolated by PCR ‐amplification, cloned and expressed in Escherichia coli , and purified by refolding from inclusion bodies. Analysis of the deduced amino acid sequence revealed high homology with members of the bacterial lipase family I.3, showing 97% identity to a putative lipase from Pseudomonas fluorescens P f0‐1, and 93% identity to a crystallized extracellular lipase from P seudomonas sp. MIS 38. A typical C‐terminal type I secretion signal and several putative Ca 2+ binding sites were also identified. Experimental data confirmed that L ip I .3 requires Ca 2+ ions for correct folding and activity. The enzyme differs from the previously reported family I .3 lipases in optimal p H , being the first acidophilic lipase reported in this family. Furthermore, L ip I .3 shows a strong preference for medium chain fatty acid esters and does not display interfacial activation. When tested for activity on secondary alcohol hydrolysis, L ip I .3 displayed higher efficiency on aromatic alcohols rather than on alkyl alcohols. Conclusions A new family I .3 lipase with unusual properties has been isolated, cloned and described. This will contribute to a better knowledge of family I .3 lipases, a family that has been scarcely explored, and that might provide a novel source of biocatalysts. Significance and Impact of the Study The unusual properties shown by L ip I .3 and the finding of activity and enantioselectivity on secondary alcohol esters may contribute to the development of new enzymatic tools for applied biocatalysis.